Simulated Cholinergic Reinnervation of β (INS-1) Cells: Antidiabetic Utility of Heterotypic Pseudoislets Containing β Cell and Cholinergic Cell

Figure 3

Coculture system hardly affects proliferation of INS-1 cells. (a, c, and d) Cellular viability was determined by CCK-8 kits. Results are means ± SD of four independent experiments. (a) INS-1 cells were cultured in complete medium (control) with or without carbachol (1 μM, 10 μM) for 48 and 72 hours. The data were expressed as percentages of untreated control cells and ns versus INS-1 control group, . (b) INS-1 cells were cultured in complete medium (control) with or without 10 μM carbachol for 72 hours. EdU-labeled INS-1 cells were assessed using fluorescent microscopy. All cell nuclei were Hoechst 33342-positive (blue), and all replicating cells were EdU-positive (red). The scatter plot represents the percentage of EdU-labeled proliferative INS-1 cells. Data were presented as the mean ± SD. ns versus control group, . (c) Cells were cultured in coculture medium (control) with or without 10 μM atropine sulfate for 48 and 72 hours. The data were expressed as 450 nm OD value and ns versus control group, . (d) Cells were cultured in coculture medium for 48 and 72 hours. The data were expressed as 450 nm OD value and ns versus the sum of INS-1 group and NG108-15 group, .

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